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干旱气候已严重影响需水作物大豆的产量,而植物中C2H2型转录因子在应答非生物逆境中起到重要作用,因此充分挖掘优异抗旱大豆种质的基因资源及功能研究,为利用基因工程手段获得抗旱大豆种质奠定理论基础。本研究从大豆叶片中克隆到2个基因,分别命名为GmZAT9-like和GmZAT5-like。序列特征分析表明二者都属于C2H2型转录因子,均包含典型双锌指结构域和EAR保守基序,且氨基酸同源性低于其他物种同源基因。分子进化树分析表明,2个基因分别与拟南芥AtZAT9和ATZAT5基因划分为一类。基于荧光实时定量PCR技术检测到2个基因分别在叶片和根部组织特异性表达。通过半定量RT-PCR和荧光定量PCR分析大豆幼苗在PEG、SA、ABA、NaCl和4℃胁迫处理条件下2个基因的表达模式。结果表明,在PEG和SA胁迫条件下,叶片中GmZAT9-like基因表达在处理后期有升高趋势,而根部GmZAT5-like基因在处理早期受到诱导表达;ABA胁迫条件下,2个基因均在处理初期呈现表达升高趋势而后下降;盐和冷胁迫条件下,叶片中GmZAT9-like基因表达受到抑制,而根部GmZAT5-like基因表达在冷胁迫处理初期呈现升高趋势。因此推测这两个基因与大豆非生物胁迫响应相关。
Arid climate has seriously affected the yield of water-crop soybean, and plant C2H2-type transcription factor plays an important role in response to abiotic stress. Therefore, fully explore the genetic resources and function of drought-tolerant soybean germplasm for genetic engineering Access to drought-resistant soybean germplasm to lay the theoretical foundation. In this study, two genes were cloned from soybean leaves and named GmZAT9-like and GmZAT5-like, respectively. Sequence analysis showed that both of them were C2H2 type transcription factors, both of which contained the typical double zinc finger domain and EAR conserved motif, and the amino acid homology was lower than that of other species homologous genes. Molecular phylogenetic tree analysis showed that the two genes were classified into Arabidopsis AtZAT9 and ATZAT5 genes respectively. Based on real-time fluorescence quantitative PCR, two genes were detected specifically in the leaf and root tissues respectively. Semi-quantitative RT-PCR and real-time quantitative PCR were used to analyze the expression patterns of two genes in soybean seedlings under PEG, SA, ABA, NaCl and 4 ℃ stress conditions. The results showed that the expression of GmZAT9-like gene in leaves increased under the PEG and SA stress conditions, whereas the root GmZAT5-like gene was induced at the early stage of treatment. Under ABA stress, the two genes were all treated The expression of GmZAT9-like gene was inhibited in salt and cold stress, while the expression of GmZAT5-like gene in root showed an increasing trend in the initial stage of cold stress treatment. Therefore, we speculated that these two genes were related to the response of soybean abiotic stress.